An automated rate analysis of lipase, trypsin, and chymotrypsin reactions by constant pH titration
✍ Scribed by A. Vandermeers; H. Lelotte; J. Christophe
- Publisher
- Elsevier Science
- Year
- 1971
- Tongue
- English
- Weight
- 477 KB
- Volume
- 42
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
An Automated Rate Analysis of Lipase, Trypsin, and Chymotrypsin Reactions by Constant pH Titration Constant pH titration methods previously used in our laboratory (1,2) for the estimation of hydrolaseh in panc'reas l~omogenatcs and in intestinal juice have been automated by an alternating stop and flow system (3). A s:tmpler and a pump provided hy Technicon Corporation ww conncctfd with an inexpensive progranmird :wseinbly of clectromagnetic valves for the automatic sulq~lp of R classical pH-stat unit. Reagerl fs Tris-HCZ bzr.fler. The buffer used for dissolving all suhstratw was matlc of 5 mM Tris-HCl (pH 8.0) rnriched with 40 m!l1 NaCl ant1 30 mJ1 CaCl,. I,iptrse substrate.
This reagent was madr daily hy adding 50 ml of the I~ffer to 30 ml of a loo/, olive oil cmulrion prepared in 10% gum acacia. The diluted emulsion was adjusted to pH 8.8 with 1 ~11 NaOH, kept' on ice. and magnetically st'irrcd during it's utilizat#ion. Trypsin substmte.